Product Pathways - PathScan ELISA
PathScan® Phospho-p70 S6 Kinase (Thr389) Chemiluminescent Sandwich ELISA Kit #7153
|7153S||1 Kit (96 assays)||---||In Stock||---|
|7153||carrier free and custom formulation / quantity||email request|
When ordering five or more kits, please contact us for processing time and pricing at firstname.lastname@example.org.
Already purchased this product? Write a Review.
|Kit Includes||Volume||Solution Color|
|p70 S6 Kinase Rabbit Antibody Coated Microwells||96 tests|
|Phospho-p70 S6 Kinase (Thr389) Mouse Detection Antibody||5.5 ml||Green|
|Anti-Mouse IgG HRP-Linked Antibody||5.5 ml||Red|
|Luminol/Enhancer Solution||3 ml||Colorless|
|Stable Peroxide Buffer||3 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
Note: 12 8-well modules – Each module is designed to break apart for 8 tests.
Storage: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
The PathScan® Phospho-p70 S6 Kinase (Thr389) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-p70 S6 kinase (Thr389) protein with a chemiluminescent readout. Chemiluminescence ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA which is offered in low volume microplates, shows increased signal and sensitivity while using a smaller sample size. A p70 S6 kinase rabbit antibody has been coated on the microwells. After incubation with cell lysates, both phospho- and nonphospho-p70 S6 kinase proteins are captured by the coated antibody. Following extensive washing, phospho-p70 S6 kinase (Thr389) mouse antibody is added to detect the captured phospho-p70 S6 kinase (Thr389) protein. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of phospho-p70 S6 kinase (Thr389) protein.
Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
PathScan® Phospho-p70 S6 Kinase (Thr389) Chemiluminescent Sandwich ELISA Kit #7153 detects endogenous levels of phospho-p70 S6 kinase (Thr389) in human and mouse cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Relationship between protein concentration of lysates from untreated and IGF-1-treated MCF7 cells and immediate light generation with chemiluminescent substrate is shown. Cells were treated with IGF-1 #3093 (100 ng/ml) and lysed after incubation at 37ºC for 20 minutes. Graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range.
p70 S6 kinase is a mitogen activated Ser/Thr protein kinase that is required for cell growth and G1 cell cycle progression (1,2). p70 S6 kinase phosphorylates the S6 protein of the 40S ribosomal subunit and is involved in translational control of 5' oligopyrimidine tract mRNAs (1). A second isoform, p85 S6 kinase, is derived from the same gene and is identical to p70 S6 kinase except for 23 extra residues at the amino terminus, which encode a nuclear localizing signal (1). Both isoforms lie on a mitogen activated signaling pathway downstream of phosphoinositide-3 kinase (PI-3K) and the target of rapamycin, FRAP/mTOR, a pathway distinct from the Ras/MAP kinase cascade (1). The activity of p70 S6 kinase is controlled by multiple phosphorylation events located within the catalytic, linker and pseudosubstrate domains (1). Phosphorylation of Thr229 in the catalytic domain and Thr389 in the linker domain are most critical for kinase function (1). Phosphorylation of Thr389, however, most closely correlates with p70 kinase activity in vivo (3). Prior phosphorylation of Thr389 is required for the action of phosphoinositide 3-dependent protein kinase 1 (PDK1) on Thr229 (4,5). Phosphorylation of this site is stimulated by growth factors such as insulin, EGF and FGF, as well as by serum and some G-protein-coupled receptor ligands, and is blocked by wortmannin, LY294002 (PI-3K inhibitor) and rapamycin (FRAP/mTOR inhibitor) (1,6,7). Ser411, Thr421 and Ser424 lie within a Ser-Pro-rich region located in the pseudosubstrate region (1). Phosphorylation at these sites is thought to activate p70 S6 kinase via relief of pseudosubstrate suppression (1,2). Another LY294002 and rapamycin sensitive phosphorylation site, Ser371, is an in vitro substrate for mTOR and correlates well with the activity of a partially rapamycin resistant mutant p70 S6 kinase (8).
- Pullen, N. and Thomas, G. (1997) FEBS Lett. 410, 78-82.
- Dufner, A. and Thomas, G. (1999) Exp. Cell Res. 253, 100-109.
- Weng, Q.P. et al. (1998) J. Biol. Chem. 273, 16621-16629.
- Pullen, N. et al. (1998) Science 279, 707-710.
- Alessi, D.R. et al. (1998) Curr. Biol. 8, 69-81.
- Polakiewicz, R.D. et al. (1998) J. Biol. Chem. 273, 23534-23541.
- Fingar, D.C. et al. (2002) Genes Dev. 16, 1472-1487.
- Saitoh, M. et al. (2002) J. Biol. Chem. 277, 20104-20112.
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
* Product-specific protocol.
- 7063 PathScan® Phospho-p70 S6 Kinase (Thr389) Sandwich ELISA Kit
- 7053 PathScan® Phospho-p70 S6 Kinase (Thr389) Sandwich ELISA Antibody Pair
- 7038 PathScan® Total p70 S6 Kinase Sandwich ELISA Kit
- 9234 Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb
- 9206 Phospho-p70 S6 Kinase (Thr389) (1A5) Mouse mAb
- 2708 p70 S6 Kinase (49D7) Rabbit mAb
- 9803 Cell Lysis Buffer (10X)
For Research Use Only. Not For Use In Diagnostic Procedures.
PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.